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Chromatography (TLC)
Learning objective: Students will learn that the different active ingredients of drug tablets
can be separated using thin layer chromatography
Vocabulary: capillary action, capillary tubes, thin layer chromatography, plates, stationary
phase, mobile phase, spotting, develop, solvent front, retention factor (Rf), analgesics, OTC,
eluent
Materials needed:
TLC plates
Crushed tablets of analgesics
Capillary micropipette
Pencils
Methanol to dissolve the tablets
mixture of ethylacetate,ethanol, and acetic acid (25:1:1 v/v/v)
developing jar
UV lamp to check the spots on the TLC plate
Background:
Chromatographic techniques are used extensively for routine analysis in chemistry.TLC can be
used to determine the purity of a substance and to analyze the composition of a mixture. It is
an inexpensive technique that gives the chemist a quick answer as to how many components
are in a mixture. A TLC plate consists of a thin layer of silica gel particles coated on a thin sheet
of aluminum. The silica is a stationary phase and an organic solvent is used as the mobile phase.
The solvent moves up the plate by capillary action.
A review of simple paper chromatography using material like different colored-markers will be
done using a video for demonstration. Thin layer chromatography will be introduced and a
short video of how it works will also be shown to the students.
Instructional procedure:
How the separation works:
The spots (different substances) move at different speeds on the TLC plate
Analgesic medications:
Activity outline:
The active ingredients of the tablets will be identified by spotting authentic standards of
the various components on a TLC plate
A solution of an unknown analgesic tablet will be spotted alongside the standards
The location of the spots from the standard and unknown will be compared
The identification of the brand name of the tablet will be accomplished by comparing
experimental results to the list below
Procedure:
Table of Results
The lesson will be concluded by letting the students do a simple measurement of the distance
travelled by their unknown and the mobile phase. They will then calculate the Rf (retention
factor). The take home message will be that Rf values are different for different components of
a substance.
Students’ understanding of the lesson will be evaluated by how well they rationalize
their choice of the unknown.